EpiJET 5-hmC Analysis Kit
EpiJET 5-hmC Analysis Kit is used for 5-hmC analysis and quantification in CCGG sequences.
Detailní popis
The Thermo Scientific EpiJET 5-hmC Analysis Kit is an efficient system for detection of 5-hydroxymethylcytosine (5-hmC) within specific DNA loci. The kit takes advantage of the T4 phage β-glucosyltransferase (T4 BGT) ability to specifically modify 5-hmC residues by adding glucose moiety to 5-hmC and the sensitivity of restriction enzyme MspI to glucosylated 5-hmC.
Features
The EpiJET 5-hmC Analysis Kit includes two advanced enzymes: T4 BGT and Epi MspI. The T4 BGT is formulated for highly specific, complete, and fast glucosylation of 5-hmC in 15 min. The Epi MspI is an MspI restriction enzyme specially formulated for efficient use in epigenetic studies. It completely digests genomic DNA in 4 hours when cytosine, 5-methylcytosine, or 5-hydroxymethylcytosine is within its recognition sequence, but its activity is completely blocked when 5-hmC is glucosylated. The analysis involves DNA sample glucosylation and subsequent Epi MspI digestion. Restriction digestion products are then analyzed by qPCR with a primer pair flanking CCGG
site of interest (see Figure 1 in Supporting Data). Use the specially designed calculator for calculation of 5-hmC percentage.
Highlights
- Complete and specific 5-hmC conversion to glucosylated 5-hmC in 15 min
- Discrimination of 5-hmC in
CCGG
sequences by Epi MspI digestion in 4 hours - Quantification of 5-hmC percentage in
CCGG
sequences by qPCR
Applications
- 5-hmC analysis and quantification in
CCGG
sequences
Includes
- T4 β – glucosyltransferase, 5 U/µL
- 10X Epi Buffer
- 10X UDP–glucose
- Epi MspI
- 5-hmC Control DNA
- Control primer 1
- Control primer 2
- Water, nuclease-free .
- Detailed Protocol
Hazardous | No |
Quality Control |
|
REviewer | Use for DNA sequence analysis and map creation. |
Storage Condition | -20 C |
Obsah balení
25 rxns
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